Evaluation of neuroprotective properties of two synthetic prenylated xanthone analogues against paraquat and 6- hydroxydopamine toxicity in human neuroblastoma SHSY5Y cells

Purpose: To investigate whether two synthetic prenylated xanthone analogues - 1,3,6,8-tetrahydroxy- 9H-xanthen-9-one (SX1) and 1,3,6-trihydroxy-2-(3-methylbut-2-enyl)-9H-xanthen-9-one (SX2) - are potential candidates for neuroprotection against paraquat- and 6-hydroxydopamine (OHDA)-induced human neuroblastoma SH-SY5Y cell death.Methods: SH-SY5Y cells were treated with SX1 and SX2, and the maximum non-toxic dose (MNTD) were obtained by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. MTT assay was also used to assess the ability of MNTD and half MNTD (HMNTD) doses of SX1 and SX2 to protect against the neurotoxicity of 200 μM paraquat and 100 μM 6-OHDA. Intracellular ROS production by SHSY5Y cells treated or untreated with SX1 or SX2 was measured by dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay.Results: SX1 and SX2 MNTDs at concentrations of 1850 nM and 105 nM, respectively, did not significantly (p > 0.05) provide neuroprotection against paraquat-induced SH-SY5Y cell death. Only SX2 MNTD and HMNTD significantly (p < 0.05) protected SH-SH5Y cells against 6-OHDA-induced cell death by 10 and 17 % improved cell viability. Although intracellular ROS production was significantly attenuated by SX1 HMNTD and MNTD, this did not improve cell viability against paraquat-induced cell death.Conclusion: These results reveal that SX2 confers neuroprotection on 6-OHDA-induced SH-SY5Y neurotoxicity. Further investigations to elucidate the detailed molecular mechanisms of neuroprotection by SX2 are warranted.Keywords: Prenylation, Xanthone derivatives, Neuroprotection, Paraquat, Dopamine, Neurotoxicity, Human neuroblastoma SH-SY5Y cell, 6-Hydroxydopamin

Anti-obesity effect of phenylcoumarins from two Calophyllum spp in 3T3-L1 adipocytes

Purpose: To evaluate the anti-obesity effects of five compounds isolated from Calophyllum andersonnii and Calophyllum sclerophyllum, viz, friedelin (CP1), friedelinol (CP2), isodispar B (CP3), 5,7-dihydroxy-6-(3-methybutyryl)-4-phenylcoumarin (CP4) and 5,7-dihydroxy-6-(2-methybutyryl)-4-phenylcoumarin (CP5) in 3T3-L1 mouse pre-adipocytes.Methods: Maximum non-toxic doses (MNTDs) of CP1 - CP5 were obtained by conducting 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Intracellular lipid droplet accumulation was determined by Oil Red O (ORO) staining. The effects of CP1 - 5 on the expression of adipogenesis transcriptional factors, namely, C/ebpα, Pparγ1, aP2 and on cellular glucose uptake and adipokine (adiponectin, leptin, resistin) secretion were assessed by commercial colorimetric and ELISA kits, respectively.Results: MNTDs for CP1-CP5 were 3.0, 1.4, 1.0, 29.0 and 25.0 μM, respectively. 3T3-L1 cells treated with CP1 - CP3 showed increased lipid accumulation (p < 0.05) and decreased glucose uptake (p < 0.05), compared with untreated cells; cells treated with CP4 and CP5 had opposite effects. Cells treated with CP4 and CP5 also showed downregulated Pparγ1, C/ebpα and aP2 expression (p < 0.05), compared with untreated cells. The anti-adipogenic property exerted by CP4 and CP5 manifested as increased secretion of adiponectin as well as reduced leptin and resistin levels.Conclusion: CP4 and CP5 isolated from Calophyllum sclerophyllum show promising anti-obesity properties, and could serve as candidate hits for further investigation at in vivo level to provide additional mechanistic evidence.Keywords: Phenylcoumarin, Calophyllum, Adipogenesis, Adipocyte, Adipokine, Obesit

A Review on Medicinal Properties of Orientin

Medicinal plants continue to play an important role in modern medications and healthcare as consumers generally believe that most of them cause fewer or milder adverse effects than the conventional modern medicines. In order to use the plants as a source of medicinal agents, the bioactive compounds are usually extracted from plants. Therefore, the extraction of bioactive compounds from medicinal plants is a crucial step in producing plant-derived drugs. One of the bioactive compounds isolable from medicinal plants, orientin, is often used in various bioactivity studies due to its extensive beneficial properties. The extraction of orientin in different medicinal plants and its medicinal properties, which include antioxidant, antiaging, antiviral, antibacterial, anti-inflammation, vasodilatation and cardioprotective, radiation protective, neuroprotective, antidepressant-like, antiadipogenesis, and antinociceptive effects, are discussed in detail in this review

Angiotensinogen M235T gene variants and its association with essential hypertension and plasma renin activity in Malaysian subjects: A case control study

BACKGROUND: Essential hypertension is a major public health concern worldwide where its prevalence accounts for various cerebrovascular diseases. A common molecular variant of angiotensinogen (AGT), the precursor of potent vasoactive hormone angiotensin II, has been incriminated as a marker for genetic predisposition to essential hypertension in some ethnics. This case-control study was designed not only to determine the association of the AGT M235T gene variants with essential hypertension, but also its relationship to Plasma Renin Activity (PRA) in subjects attending the Health Clinic, Kuala Lumpur, Malaysia. METHODS: The study involved 188 subjects, 101 hypertensives and 87 normotensives. Consents were obtained from all the participated subjects. M235T gene variants were investigated using allele specific polymerase chain reaction and PRA was determined by radioimmunoassay. Hypertensinogenic factors such as dietary habits, physical activity, smoking and drinking habits were assessed using a pre-tested questionnaire. RESULTS: The genotype and allele distribution of the M235T variant differed significantly in hypertensives and normotensives (χ(2 = )23.184, P < 0.001 and χ(2 )= 21.482, P < 0.001, respectively). The odds ratio for hypertension was 1.36 (95% confidence interval 1.03–1.80) for subjects with homozygous mutated allele TT of the M235T variant compared with other genotypes or 1.98 (95% confidence interval 1.46–2.67) for those carrying T allele compared to those carrying M allele. Plasma Renin Activity is also significantly higher in hypertensive subjects (PRA = 3.8 ± 2.5 ngAI/ml/hr for hypertensives, PRA = 2.6 ± 1.3 ngAI/ml/hr for normotensives, P < 0.001), but was not significantly different between groups of genotypes (P = 0.118). CONCLUSION: The M235T variant of the AGT is significantly associated with essential hypertension whereas the genotype TT or allele T is a possible genetic marker or risk factor for hypertension in Malaysian subjects

The association of insertions/deletions (INDELs) and variable number tandem repeats (VNTRs) with obesity and its related traits and complications

Abstract Background Despite the fact that insertions/deletions (INDELs) are the second most common type of genetic variations and variable number tandem repeats (VNTRs) represent a large portion of the human genome, they have received far less attention than single nucleotide polymorphisms (SNPs) and larger forms of structural variation like copy number variations (CNVs), especially in genome-wide association studies (GWAS) of complex diseases like polygenic obesity. This is exemplified by the vast amount of review papers on the role of SNPs and CNVs in obesity, its related traits (like anthropometric measurements, biochemical variables, and eating behavior), and its related complications (like hypertension, hypertriglyceridemia, hypercholesterolemia, and insulin resistance—collectively known as metabolic syndrome). Hence, this paper reviews the types of INDELs and VNTRs that have been studied for association with obesity and its related traits and complications. Main body of the abstract These INDELs and VNTRs could be found in the obesity loci or genes from the earliest GWAS and candidate gene association studies, like FTO, genes in the leptin–proopiomelanocortin pathway, and UCP2/3. Given the important role of the brain serotonergic and dopaminergic reward system in obesity susceptibility, the association of INDELs and VNTRs in these neurotransmitters’ metabolism and transport genes with obesity is also reviewed. Next, the role of INS VNTR in obesity and its related traits is questionable, since recent large-scale studies failed to replicate the earlier positive associations. As obesity results in chronic low-grade inflammation of the adipose tissue, the proinflammatory cytokine gene IL1RA and anti-inflammatory cytokine gene IL4 have VNTRs that are implicated in obesity. A systemic proinflammatory state in combination with activation of the renin–angiotensin system and decreased nitric oxide bioavailability as found in obesity leads to endothelial dysfunction. This explains why VNTR and INDEL in eNOS and ACE, respectively, could be predisposing factors of obesity. Finally, two novel genes, DOCK5 and PER3, which are involved in the regulation of the Akt/MAPK pathway and circadian rhythm, respectively, have VNTRs and INDEL that might be associated with obesity. Short conclusion In conclusion, INDELs and VNTRs could have important functional consequences in the pathophysiology of obesity, and research on them should be continued to facilitate obesity prediction, prevention, and treatment

Palmitic acid induces neurotoxicity and gliatoxicity in SH-SY5Y human neuroblastoma and T98G human glioblastoma cells

Background Obesity-related central nervous system (CNS) pathologies like neuroinflammation and reactive gliosis are associated with high-fat diet (HFD) related elevation of saturated fatty acids like palmitic acid (PA) in neurons and astrocytes of the brain. Methods Human neuroblastoma cells SH-SY5Y (as a neuronal model) and human glioblastoma cells T98G (as an astrocytic model), were treated with 100–500 µM PA, oleic acid (OA) or lauric acid (LA) for 24 h or 48 h, and their cell viability was assessed by 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of stable overexpression of γ-synuclein (γ-syn), a neuronal protein recently recognized as a novel regulator of lipid handling in adipocytes, and transient overexpression of Parkinson’s disease (PD) α-synuclein [α-syn; wild-type (wt) and its pathogenic mutants A53T, A30P and E46K] in SH-SY5Y and T98G cells, were also evaluated. The effects of co-treatment of PA with paraquat (PQ), a Parkinsonian pesticide, and leptin, a hormone involved in the brain-adipose axis, were also assessed. Cell death mode and cell cycle were analyzed by Annexin V/PI flow cytometry. Reactive oxygen species (ROS) level was determined using 2′,7′-dichlorofluorescien diacetate (DCFH-DA) assay and lipid peroxidation level was determined using thiobarbituric acid reactive substances (TBARS) assay. Results MTT assay revealed dose- and time-dependent PA cytotoxicity on SH-SY5Y and T98G cells, but not OA and LA. The cytotoxicity was significantly lower in SH-SY5Y-γ-syn cells, while transient overexpression of wt α-syn or its PD mutants (A30P and E46K, but not A53T) modestly (but still significantly) rescued the cytotoxicity of PA in SH-SY5Y and T98G cells. Co-treatment of increasing concentrations of PQ exacerbated PA’s neurotoxicity. Pre-treatment of leptin, an anti-apoptotic adipokine, did not successfully rescue SH-SY5Y cells from PA-induced cytotoxicity—suggesting a mechanism of PA-induced leptin resistance. Annexin V/PI flow cytometry analysis revealed PA-induced increase in percentages of cells in annexin V-positive/PI-negative quadrant (early apoptosis) and subG0-G1 fraction, accompanied by a decrease in G2-M phase cells. The PA-induced ROS production and lipid peroxidation was at greater extent in T98G as compared to that in SH-SY5Y. Discussion In conclusion, PA induces apoptosis by increasing oxidative stress in neurons and astrocytes. Taken together, the results suggest that HFD may cause neuronal and astrocytic damage, which indirectly proposes that CNS pathologies involving neuroinflammation and reactive gliosis could be prevented via the diet regimen

PA SHT98 Paper Raw Data YHSay

Raw data for the paper <b>"Palmitic acid induces neurotoxicity and gliatoxicity in SH-SY5Y human neuroblastoma and T98G human glioblastoma cells" </b

Association of fat mass and obesity-associated (FTO) gene rs9939609 variant with obesity among multi-ethnic Malaysians in Kampar, Perak

Obesity is a multifactorial disease caused by the interaction of genetic, lifestyle and environmental factors. Common single nucleotide polymorphisms in the recently-described Fat Mass and Obesity-Associated (FTO) gene have been related to body weight and fat mass in humans and genome-wide association studies in several populations have indicated that the FTO rs9939609 variant is associated with obesity. Therefore, the objective of this study was to investigate the association of the FTO rs9939609 variant with obesity among 324 multi-ethnic Malaysians (98 Malays, 158 Chinese, 68 Indians) who attended the Kampar Health Clinic, Perak. With the overall minor A allele frequency (MAF) of 0.199, the distribution of genotypes and alleles was significantly different among ethnicities (MAF highest among Malays), but no association was found for obesity, related anthropometric measurements and gender. Subject with allele A had marginally but significantly higher waist circumference (p=0.015), thus the FTO rs9939609 allele was associated with central obesity (p=0.034 by Chi-square analysis; Odds Ratio (OR)=1.680 (CI=1.036, 2.724; p=0.035)). However, this association was abolished when adjusted for age, gender and ethnicity (OR=1.455, CI=0.874, 2.42; p=0.149). In conclusion, the MAF of the FTO rs9939609 SNP was low as in other Asian populations and there was no evidence for an involvement of this SNP in obesity and obesity-related traits in this multi-ethnic Malaysian study group

Gender Differences in Body Image Perception among Northern Malaysian Tertiary Students

Aims: This study examined the association of socio-cultural and psychological factors with body shape concern, perception and body weight perception among tertiary students of Northern Malaysia. Study Design: This is a cross-sectional study. Place and Duration of Study: Universiti and Kolej Tunku Abdul Rahman (UTAR and KTAR), Perak campuses, between August 2011 and January 2012. Methodology: A total of 1003 students were recruited (M = 431, F = 572; mean age 19.96 ± 1.51) and their body image perception were assessed using Body Shape Concern Questionnaire, Body Weight Perception Questionnaire, Body Shape Perception Questionnaire (Stunkard Silhouette Chart), Multidimensional Body Self Relations Questionnaire (MBSRQ), Rosenberg’s Self-Esteem Scale (RSE) and Quality of Life measurement. Results: More females than males had problems with their body shape, where more females desired a thinner body size and vice versa for males. There was misperception of opposite sex’s perception of attractive body shape, where males chose a larger figure for attractive body shape of female compared to females themselves, and vice versa. Overweight students had significantly lower parental/peer acceptance, higher body shape satisfaction and hence lower body weight/shape anxiety, and made lesser body shape comparison compared to other counterparts. Quality of life and self-esteem were significantly negatively correlated with body satisfaction. Conclusion: Male and female Malaysian tertiary students were concerned with their body shape and perceived their body weight/shape differently